May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Evaluation of in vitro Effects of Triamcinolone Acetonide and Dexamethasone on Human Lens Epithelial Cells
Author Affiliations & Notes
  • A. Pirouzmanesh
    Ophthalmology, Univ of California - Irvine, Irvine, California
  • A. Sharma
    Ophthalmology, Univ of California - Irvine, Irvine, California
  • A. Pirouzmanesh
    Ophthalmology, Univ of California - Irvine, Irvine, California
  • U. P. Andley
    Department of Ophthalmology and Visual Sciences,, Washington University, St. Louis, Missouri
  • M. C. Kenney
    Ophthalmology, Univ of California - Irvine, Irvine, California
  • B. D. Kuppermann
    Ophthalmology, Univ of California - Irvine, Irvine, California
  • Footnotes
    Commercial Relationships  A. Pirouzmanesh, None; A. Sharma, None; A. Pirouzmanesh, None; U.P. Andley, None; M.C. Kenney, None; B.D. Kuppermann, None.
  • Footnotes
    Support  Supported by the Discovery Eye Foundation, the Henry L. Guenther Foundation, the Iris and B. Gerald Cantor Foundation, Gilbert Foundation and Research to Prevent Blindness Foundation.
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 2776. doi:https://doi.org/
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      A. Pirouzmanesh, A. Sharma, A. Pirouzmanesh, U. P. Andley, M. C. Kenney, B. D. Kuppermann; Evaluation of in vitro Effects of Triamcinolone Acetonide and Dexamethasone on Human Lens Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2008;49(13):2776. doi: https://doi.org/.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To compare the toxicity of triamcinolone acetonide (TA) and dexamethasone sodium phosphate (DEX) on human lens epithelial (HLE) cells in-vitro.

Methods: : HLE cells were cultured in Minimum Essential Medium (MEM). Commercially available TA [(c-TA) (Kenalog®, BMS, Princeton, NJ)] was centrifuged at 5000 rpm for one minute and the supernatant was removed. The pellet was re-suspended in equivalent amounts of DMSO to produce solubulized TA (s-TA). Cells were treated with 100, 200, 500, 750, or 1000 (clinical dose) µg/mL concentrations of c-TA, s-TA, and the supernatant for 24 hours. Other HLE cells were treated with dexamethasone sodium phosphate (APP, Schaumburg, IL) at 0.05, 0.1 (clinical dose), 0.2, 0.5, 1, and 2 mg/ml. Cell viability was determined by trypan blue dye-exclusion assay. Caspase-3/7 was measured by fluorescence caspase kit.

Results: : The mean cell viabilities of HLE after 24 hours exposure to c-TA at 1000, 750, 500, 200, and 100µg/ml were 17.3 ± 5.5 (P < 0.001), 23.2 ± 6.2 (P < 0.001), 31.8 ± 6.8 (P < 0.001), 52.9 ± 5.9 (P <0.01), and 71.1 ± 1.65 (P <0.05), respectively, compared to control untreated cells. The mean cell viabilities of HLE after exposure to s-TA at 1000, 750, 500, 200, and 100µg/ml were 19.8 ± 1.1 (P < 0.001), 28.6 ± 3.2 (P < 0.001), 38.4 ± 2.4 (P < 0.001), 77.3 ± 3.1 (P >0.05), and 80.5 ± 2.45 (P >0.05), respectively, compared to equivalent DMSO control. The supernatant alone did not reduce the viability of HLE. Caspase-3/7 activity in HLE cells increased significantly after treatment with s-TA at 750, and 1000 µg/mL.The mean cell viabilities of HLE after 24 hours exposure to DEX at 2 , 1, 0.5, 0.2, 0.1, and 0.05 mg/ml were 14.8 ± 0.6 (P < 0.001), 23.6 ± 0.1 (P < 0.001), 80.1 ± 0.9 (P < 0.001), 92.4 ± 0.3 (P > 0.05), 93.1 ± 2.1 (P > 0.05), and 93.7 ± 2.1 (P > 0.05), respectively, compared to control untreated cells. Caspase-3/7 activities in HLE cells were not increased significantly after treatment with DEX at all concentrations tested.

Conclusions: : This study showed that TA at its clinical dose in both commercial preparation and solubilized form decreases human lens epithelial cell viability and that the caspase-3/7 pathway is involved suggestive of an apoptotic pathway. DEX at its clinical dose does not decrease cell viability nor cause any increase of capase-3/7 activity.

Keywords: drug toxicity/drug effects • corticosteroids • apoptosis/cell death 
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